3N5F

Crystal Structure of L-N-carbamoylase from Geobacillus stearothermophilus CECT43

3N5F の概要

• エントリーDOI: 10.2210/pdb3n5f/pdb
• 分子名称: N-carbamoyl-L-amino acid hydrolase, ISOPROPYL ALCOHOL, COBALT (II) ION, ... (5 entities in total)
• 機能のキーワード: carbamoylase, hinge domain, m20 peptidase family, evolution, binding residue, dimerization domain, hydrolase
• 由来する生物種: Bacillus stearothermophilus (Geobacillus stearothermophilus)
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 88715.67

構造登録者

Garcia-Pino, A.,Martinez-Rodriguez, S.,Gavira, J.A. (登録日: 2010-05-25, 公開日: 2011-05-25, 最終更新日: 2023-09-06)

主引用文献

Martinez-Rodriguez, S.,Garcia-Pino, A.,Las Heras-Vazquez, F.J.,Clemente-Jimenez, J.M.,Rodriguez-Vico, F.,Garcia-Ruiz, J.M.,Loris, R.,Gavira, J.A.
Mutational and structural analysis of L-N-carbamoylase reveals new insights into a peptidase m20/m25/m40 family member.
J.Bacteriol., 194:5759-5768, 2012

PubMed Abstract: N-Carbamoyl-L-amino acid amidohydrolases (L-carbamoylases) are important industrial enzymes used in kinetic resolution of racemic mixtures of N-carbamoyl-amino acids due to their strict enantiospecificity. In this work, we report the first L-carbamoylase structure belonging to Geobacillus stearothermophilus CECT43 (BsLcar), at a resolution of 2.7 Å. Structural analysis of BsLcar and several members of the peptidase M20/M25/M40 family confirmed the expected conserved residues at the active site in this family, and site-directed mutagenesis revealed their relevance to substrate binding. We also found an unexpectedly conserved arginine residue (Arg(234) in BsLcar), proven to be critical for dimerization of the enzyme. The mutation of this sole residue resulted in a total loss of activity and prevented the formation of the dimer in BsLcar. Comparative studies revealed that the dimerization domain of the peptidase M20/M25/M40 family is a "small-molecule binding domain," allowing further evolutionary considerations for this enzyme family.

PubMed: 22904279
DOI: 10.1128/JB.01056-12

実験手法

X-RAY DIFFRACTION (2.75 Å)